Semicontinuous Flow Electroporation Chip for High-Throughput Transfection on Mammalian Cells

Wang, Shengnian; Zhang, Xulang; Wang, Weixiong; Lee, L. James

doi:10.1021/ac9002672

Cited by 58 publications

(53 citation statements)

References 47 publications

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“…(For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.) by directed transfection [176], or directly by electrofusion [177][178][179]. For example, Schoeman et al [179] engineered a device capable of encapsulating single cells inside droplets generated in parallel, which are paired in a second step and merged by the action of an electrical field.…”

Section: Biomaterials For Cell Selection and Enrichmentmentioning

confidence: 99%

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

2016

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Microfluidics, being a technology characterized by the engineered manipulation of fluids at the submillimeter scale, offers some interesting tools that can advance biomedical research and development. Screening platforms based on microfluidic technologies that allow high-throughput and combinatorial screening may lead to breakthrough discoveries not only in basic research but also relevant to clinical application. This is further strengthened by the fact that reliability of such screens may improve, since microfluidic systems allow close mimicking of physiological conditions. Finally, microfluidic systems are also very promising as micro factories of a new generation of natural or synthetic biomaterials and constructs, with finely controlled properties.

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Section: Biomaterials For Cell Selection and Enrichmentmentioning

confidence: 99%

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

2016

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“…Nevertheless, there are efforts to achieve high throughput in MEP and NEP using array-type and flow-through setups. [65][66][67]71,[73][74][75][76][77][78][79][80][81][82][83][84][85] In these devices, either electric pulses or a DC field is imposed on the cell solution occupied the entire flow channel, or as solution droplets containing multiple cells. These technologies are briefly summarized here.…”

Section: E High Throughput For Large Scale Cell Transfectionmentioning

confidence: 99%

“…Better transgene expression (10%-15% higher) of pWizGFP plasmid and a 50% or higher cell viability on K562 cells and mouse embryonic stem cells were achieved. 84 To further minimize the electrolysis issue, a similar flow-through electroporation setup integrated with hydrodynamic focusing flow pattern was recently tested by Wei et al (Fig. 5(f)).…”

Section: E High Throughput For Large Scale Cell Transfectionmentioning

confidence: 99%

Micro-/nanofluidics based cell electroporation

Wang

Lee

2013

Biomicrofluidics

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Non-viral gene delivery has been extensively explored as the replacement for viral systems. Among various non-viral approaches, electroporation has gained increasing attention because of its easy operation and no restrictions on probe or cell type. Several effective systems are now available on the market with reasonably good gene delivery performance. To facilitate broader biological and medical applications, micro-/nanofluidics based technologies were introduced in cell electroporation during the past two decades and their advances are summarized in this perspective. Compared to the commercially available bulk electroporation systems, they offer several advantages, namely, (1) sufficiently high pulse strength generated by a very low potential difference, (2) conveniently concentrating, trapping, and regulating the position and concentration of cells and probes, (3) real-time monitoring the intracellular trafficking at single cell level, and (4) flexibility on cells to be transfected (from single cell to large scale cell population). Some of the micro-devices focus on cell lysis or fusion as well as the analysis of cellular properties or intracellular contents, while others are designed for gene transfection. The uptake of small molecules (e.g., dyes), DNA plasmids, interfering RNAs, and nanoparticles has been broadly examined on different types of mammalian cells, yeast, and bacteria. A great deal of progress has been made with a variety of new micro-/nanofluidic designs to address challenges such as electrochemical reactions including water electrolysis, gas bubble formation, waste of expensive reagents, poor cell viability, low transfection efficacy, higher throughput, and control of transfection dosage and uniformity. Future research needs required to advance micro-/nanofluidics based cell electroporation for broad life science and medical applications are discussed.

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“…The available protocols are established by trial-and-error, instead of equation (1), at a compromise of acceptable transfection efficiency and cell viability. The high-voltage pulses, though effective in improving the cell membrane permeability and probe uptake, inevitably leads to severe side effects detrimental to later cell survival232425.…”

mentioning

confidence: 99%

“…500–1000 V/cm) for successful cell membrane breakdown. These microelectroporation systems open new routes towards the elimination of aforementioned electroporation induced apoptosis and simultaneously offer some other advantages over the commercial systems, namely in situ monitoring of intracellular content transport and electroporation dynamics at single cell level3940414243, better accuracy, and flexibility on treatment for different cell populations44454647484950515253. However, most of these microelectroporation systems still ignore the variations among individual cells of a large population, leaving many factors still uncontrolled just like in those commercial systems.…”

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confidence: 99%

Size Specific Transfection to Mammalian Cells by Micropillar Array Electroporation

Huang

et al. 2016

Sci Rep

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Electroporation serves as a promising non-viral gene delivery approach, while its current configuration carries several drawbacks associated with high-voltage electrical pulses and heterogeneous treatment on individual cells. Here we developed a new micropillar array electroporation (MAE) platform to advance the electroporation-based delivery of DNA and RNA probes into mammalian cells. By introducing well-patterned micropillar array texture on the electrode surface, the number of pillars each cell faces varies with its plasma membrane surface area, despite their large population and random locations. In this way, cell size specific electroporation is conveniently carried out, contributing to a 2.5~3 fold increase on plasmid DNA transfection and an additional 10–55% transgene knockdown with siRNA probes, respectively. The delivery efficiency varies with the number and size of micropillars as well as their pattern density. As MAE works like many single cell electroporation are carried out in parallel, the electrophysiology response of individual cells is representative, which has potentials to facilitate the tedious, cell-specific protocol screening process in current bulk electroporation (i.e., electroporation to a large population of cells). Its success might promote the wide adoption of electroporation as a safe and effective non-viral gene delivery approach needed in many biological research and clinical treatments.

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Semicontinuous Flow Electroporation Chip for High-Throughput Transfection on Mammalian Cells

Cited by 58 publications

References 47 publications

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

High-throughput screening approaches and combinatorial development of biomaterials using microfluidics

Micro-/nanofluidics based cell electroporation

Size Specific Transfection to Mammalian Cells by Micropillar Array Electroporation

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